5, 6-dichloro pregnanes



United States Patent() 2,884,417 5,6-DICHLORO PREGNANES Frank A. Cutler, Jr., and John M. Chemerda, Metuchen, NJ., assignors to Merck & '00., Inc., Rahway, NJ., a corporation of New Jersey Application December 17, 1952 Serial No. 326,576

5 Claims. (Cl. 260-23955) No Drawing.

novel steroid compounds and (imom i -011 Ice Patented Apr. 28, 1959 The discovery of the remarkable effectiveness of cortisone and similar related compounds in the treatment of various diseases has stimulated great interest infinding other methods of preparing these compounds.

It is an ob ect of the present invention to provide a process for the preparation of A -3,20-diketo-17a,2l-dihydroxy-pregnene (Reichsteins Substance S), an active adrenal hormone and 13 -3,2O-diketo-17a-hydroxy-pregnene. It is a further object to provide novel derivatives of pregnenolone which are useful as intermediates in the synthesis of these products. Another object is to provide processes for the preparation of said intermediates. Other objects will be apparent from the detailed description of our invention hereinafter provided.

In accordance with the present invention, we have found that A -3,20-diketo-l7d,21-dihydroxyepregnene can =0 iii-on.

43-03 l--- on onion. 03,03, :0 42 0 Li -on i orr be prepared from pregnenolone pregnene) by a process which can be represented as follows:

. (VIII) (A -3-hydroxy-20-keto- CHgOH wherein R represents an acyl substituent, and R and R; represent lower aliphatic acid radicals.

In this process, an acyl derivative of pregnenolone (I) is first chlorinated to saturate the A -unsaturated group and obtain the corresponding dichloro derivative (II) to produce the corresponding A Upon reacting the enol acylate with an OXlCllZlIlg agent the corresponding 17,20- epoxide (IV) is obtained which on hydrolysis is converted to the 17a-hydroxy-20-keto compound (V). The latter compound is then reacted with bromine to produce the corresponding 2l-bromo compound (VI) which is converted to the 21-acyloxy compound (VII) by treatment with a salt of a lower aliphatic acid. Upon treating VII 3-keto com- After completion of the reac t1on, the solution is washed successively with dilute evaporated by warming under diminished pressure to obtain the'desired 5, -dichloro compound which purified by recrystallization from suitable solvents.

The second step of our process the 5,6-dichloro compound with an excess of lower aliphatic acid one to six hours.

reactants at a temperature of about 60-110 C. for-about Strong acids, such as concentrated sulfuric acid, benzene sulfonic acid, p-toluene sulfonic for this reaction.

dichloropregnene is conveniently obtained by heating 3-acetoxy- 5,6-dichloro-20-keto-pregnane with acetic anhydride in the presence of p-toluene sulfonic acid on a steam bath for about four to five hours. After completion of the reaction, the product is recovered by decomposing the exbenzene,

and the like, and evaporating the The oxidation of the enol acylate (III) to the corresponding epoxy compound (IV) is effected by allowing the resulting reaction mixture to warm up to room temperature, and permitting the reaction to continue at room temperature until the oxidation is substantially complete. At this point the mixture is washed with alkali to remove the excess perbenzoic acid,

For example, 20 keto 21 bromopregnane is obtained by adding a solution of bromine in chloroform to a chloroform solution of .3,17-dihydroxy-S,6-dichloro-20-keto-pregnane and maintaining the reaction mixture at about 4055 C. After completion of the reaction, the reaction mixture is washed with an aqueous solution of sodium bicarbonate, and the product is obtained in crystalline form by evaporating the chloroform solution to a small volume.

The conversion of the Zl-bromo compound (VI) to the 2l-acyloxy compound (VII) can be accomplished by several methods. The reaction can be carried out by direct metathesis with an alkali metal salt of a lower aliphatic acid. This reaction is conveniently accomplished by heating a solution of the 21-bromo compound in a suitable solvent such as acetone, with the alkali metal salt for about 4-15 hours. The alkali metal salt can be added to the reaction mixture directly or formed in situ by the reaction of equivalent amounts of the organic carboxylic acid and an alkali metal hydroxide or carbonate.

Alternatively, the 21-bromo compound can be first converted to the 21-iodo compound by treatment with an iodide salt, for example sodium iodide, and the resulting iode compound converted by metathesis as described above, to the desired Zl-acyloxy compound.

Further, as will be apparent to those skilled in the art, the Zl-acyloxy compound is also conveniently obtained by reacting the iodide salt and the alkali metal carboxylic acid salt concurrently with the 2l-bromo compound. This method is generally prefer-red in carrying out this step of our process. Thus, 3,17a-dihydroxy- 5,6-dichloro-20-keto-2l-acetoxy-pregnane is conveniently prepared by refluxing a mixture of 3,17 a-dihydroxy-5,6- dichloro-20-keto-2l-bromo-pregnane, acetone, potassium acetate, acetic acid and sodium iodide for about 4-5 hours. After adding water to the resulting reaction mixture, and distilling ofi the acetone, the desired 2l-acetoxy compound can be recovered in crystalline form.

The oxidation of the 3-hydroxyl group of compound VII to the 3-keto group of VIII is efiected by treatment with suitable strong oxidizing agents. This reaction is most conveniently effected by treatment with chromium trioxide in an aqueous solution of acetic and sulfuric acids at a temperature of about -20" C. For example, 3,20- diketo 5,6 dichloro-l7whydroxy-2l-acetoxy-pregnane is obtained by intimately contacting 3,l7m-dihydroxy-5,6- dichloro-20-keto-2l-acetoxy-pregnane with chromium trioxide in an acetic acid-sulfuric acid reaction medium. The recovery of the oxidized product is efiected by extraction of the product from the reaction mixture with a suitable solvent such as chloroform, carbon tetrachloride, benzene and the like.

The removal of chlorine from compound VIII to produce the corresponding ester of A -3,20-diketo-l7a,21- dihydroxy-pregnene is effected by treating VIII with a dehalogenating agent. Thus, this dehalogenation is readily accomplished by heating VIII with zinc or chromous chloride. For example, A -3,ZO-diketo-17a-hydroxy-21- acetoxy-pregnene is produced by intimately contacting 3,20 diketo 5,6 dichloro 17a hydroxy 21 acetoxy-pregnane with zinc in an acetic acid medium at a temperature of about 40-80 C. After the reaction is complete, the reaction mixture is filtered and the desired product is recovered in crystalline form by adding water to the resulting filtrate and cooling the aqueous solution. The esters of A -3,20-diketo-l7a,2l-dihydroxy-pregnene so obtained can be hydrolyzed by treatment with alkali to obtain Reichsteins Substance S.

In accordance with a further embodiment of the present invention, it is found that A -3,20-diketo-l7a-hydroxypregnene is produced by oxidizing 3-hydroxy,-5,6-dichlorol7u-hydroXy-ZO-ketoregnane to obtain the corresponding 3-keto compound, and dehalogenating the latter can pound. This process can be shown as follows:

CH3 CH =0 =0 l:----orr l--0rr r 0- 0:

( (XII) The oxidation of V to obtain the 3-keto compound is efiected by reacting V with a strong oxidizing agent. Thus, this reaction is conveniently effected by reacting 3 hydroxy 5,6 dichloro-17a-hydroxy-ZO-keto-pregnane with chromium trioxide in the presence of acetic acid and sulfuric acid. The 3-keto compound is recovered by adding water to the reaction mixture, extracting the resulting aqueous solution with a suitable solvent such as chloroform, and evaporating the solvent extracts.

The dehalogenation of XI to produce A -3,20-diketo- 17a-hydroxy-pregnene can be accomplished as described above for converting compound VIII to compound IX. Thus, the reaction is effected by treating 3,20-diketo-5,6-. dichloro-l7a-hydroxy-pregnane in acetone solution with chromous chloride. The dehalogenated product is recovered in crystalline form by concentrating the resulting reaction mixture under diminished pressure until the product precipitates and filtering the resulting concentrate.

In the step of removing the chloro substituents from 3,20-diketo-5,6-dichloro-l7a,2l-dihydroxy-pregnane and 3,20-diketo-5,6-dichloro-l7u-hydroxy-pregnane, the initial reaction product is believed to be the A compound which readily isomerizes to form the corresponding A compound. This isomerization is conveniently efiected by treating the reaction product with a small amount of an acid such as sulfuric acid.

The products, A -3,20-diketo-l7a,2l-dihydroxy-pregnene, the esters of A -3,ZO-diketo-l7a,2l-dihydroxy-pregnene, and A -3,2O-diketo-l7a-hydroxy-pregnene which are obtained in accordance with the processes of the present invention can be treated by microbiological fermentation processes to introduce an oxygen containing substituent at C-ll. Thus, these compounds are useful intermediates in the preparation of cortisone, and hydrocortisone, and similar related compounds.

It was indeed unexpected to find that the 5,6-double bond could be protected by the addition of chlorine in the process of our invention. Other attempts to protect this double bond by the addition of hydrogen chloride or the addition of bromine were unsuccessful. The addition of I-ICl caused some epirnerization at 0-17, and the derivative so obtained would have eliminated hydrogen chloride under the reaction conditions of our process.

Similarly, when the double bond is protected by thecarrying out the processes of the present invention.

EXAMPLE 1 was refluxed 3.5 hours and then chilled slowly, with seeding, finally chilling in ice. The crystals of pregnenolone acetate were collected, washed with cold methanol and dried at room temperature.

ing the temperature below 20 then concentrated under reduced EXAMPLE 2 3-acetoxy-5,6-dichloro-20-keto-pregnane To a solution of 25 g. of pregnenolone acetate in 940 ml. of chloroform was added solution, tubes, was cooled to 60 solution of chlorine in to 15 C. with stirring and with dissolution of the precipitate to give a colorless solution.

The solution was EXAMPLE 3 A -3,20-diacetxy-5,6-dichloro-pregnene to 10 C. and water (2235 ml.) at 10 C. was added rapidly to hydrolyze the excess acetic anhydride. The mixture was agitated and the temperature was kept below 30 C. by occasional ml.). The combined ether Water (3 x 800 ml.), 10% sodium carbonate solution (2 x 320 ml.) and water (3 x 800 ml.). The ether solution was dried over sodium sulfate and concentrated under reduced pressure.

extracts were washed with 3,20-diacetoxy-5,6-dichloro-pregnene which was used directly in the next step.

8 EXAMPLE 4 3,20-diacetoxy-5,6-dichl0r0-17,20-ep0xy-pregnane To the A -3,20 diacetoxy 5,6 -dichloro-pregnene (0.248 mole) was added 910 ml. (1.5 x 0.248 mole) of perbenzoic acid in benzene solution at 10 C. The solution was allowed to warm to room temperature and was titrated periodically. After 4.75 hours 88% of the (3 x 1000 ml.), to neutrality with water (3 x 1000 ml.), Was dried over sodium sulfate and concentrated to dryness under reduced preswas flushed with methanol, becoming crystalline. This residue was boiled with methanol ml.) but did not dissolve. The suspen- EXAMPLE 5 3,20-diacet0xy-5,6-dichl0ro-1 7,20-ep0xy-pregnane To 0.139 gram-mole of A -3,20-diacetoxy-5,6-di chloro-pregnene (previously flushed with ethyl acetate) was added 600 ml. of a 0.487 molar solution of monoin ethyl acetate (cold). The solid and the solution warmed itself to 30 C. The reaction was largely complete in three hours but was allowed to stand 2.5 days. The solution was washed free of peracid with cold 0.5 N sodium hydroxide (3 x 600 ml.) and to neutrality with Water (3 x 600 ml.), was dried over sodium sulfate and concentrated to dryness under reduced pressure. The residue was slurried with boiling methanol (70 ml.) and then chilled and filtered. The crystals were washed with cold methanol and dried, yielding 3,20-diacetoxy-5,6-dichloro-17,20-

epoxy-pregnane.

EXAMPLE 6 3,1 7a-di/zydr0xy-5,6-dich Zora-2 O-ketO-pregnane A solution of potassium hydroxide (96.3 g., 1.72 moles) in water ml.) plus methanol (717 ml.) was prepared and cooled to 25 C. A solution of acetoxy-5,6-dichloro-17,20-epoxy-pregnane (84 g., 0.172

uct was found to melt at 203204 C. dec.

EXAMPLE 7 3,1 7 a-dihydroxy-S 6 -dich Zora-2 orketo-pregnane Five grams of 3,20-diacetoxy-5,6-dichloro-17,20-epoxypregnane obtained in Example 5 was dissolved in boiling methanol (202 ml.). A solution of potassium bicarbonate (6.15 g.) in water (20.2 ml.) was added and the mixture was refluxed for three hours. Water was added, and the methanol was removed under reduced pressure, crystallization occurring in the process. The resulting aqueous suspension was chilled and filtered, and the crystals were washed with water and dried to give 3,171 dihydroxy-5,6-dichloro-20-keto-pregnane, melting at 192 196 C. (dec.). 1 EXAMPLE 8 3,17a-dihydr0xy-5,6-dichloro-20-keto-21-br0mopregnane 3,17a-dihydroxy-5,6-dichloro-20-lreto-pregnane (4.03 3.,-

10 mM.) was dissolvedin warm chloroform (120 ml.). A solution of bromine (11 mM.) in chloroform (28.1 ml. total volume) was added over a period of one hour maintaining the reaction at 46-48 C. The solution was washed with 10% sodium bicarbonate solution (50 ml.) and water (50 ml.). The chloroform solution was concentrated under reduced pressure and the residue became crystalline on warming in a small amount of methanol. After chilling, the crystals of 3,17a-dihYd1OXY-5,6- dichloro 20 keto 21 bromo pregnane were collected, washed with a little cold methanol, and dried. The

product melted with decomposition at 179.5-l81.5 C.

EXAMPLE 9 3,17a-dihydroxy-5,6-dichlor-20-keto-21 -acetoxypregnane EXAMPLE 3,1 7 a-dihydroxy-S ,6-dichloro-20-ket0-21 -acetoxypregnane A mixture of 0.32 g. of 3,17u-dihydroxy-5,6-dichloro- 20-keto-2l-bromo-pregnane, 1.0 g. of potassium acetate and 25 cc. of acetone was refluxed for four hours. Water (50 ml.) was added and the acetone was removed under reduced pressure. The resulting crystalline suspension was chilled and filtered and the crystals were washed with water and dried. When dry the product melted at 188- 192 C. (dec.). Its infrared spectrum was the same as that of an authentic specimen of 3,l7a-dihydroxy-5,6-dichloro-20-keto-2l-acetoxy-pregnane.

EXAMPLE 1 1 3,20-diketo-5,6-dichloro-17a-hydroxy-21-acetoxypregnane One gram of 3,17a-dihydroxy-S,6-dichloro-20-keto-2lacetoxy-pregnane was dissolved in acetic acid (20 ml.) and water (2 ml.) was added to prevent freezing. The solution was chilled in an ice bath. A solution of chromium trioxide (0.216 g.) in water (0.2 ml.) plus acetic acid (2.5 ml.) was added over ten minutes while stirring in the ice bath. Sulfuric acid (conc., 0.121 ml.) was then added over two minutes as stirring continued, in the ice bath. After 20 minutes a heavy precipitate had formed. After 70 minutes further, the mixture was shaken with chloroform (50 ml.), and water (50 ml.). The chloroform layer was washed with 2.5% sodium bicarbonate solution (3 x 50 ml.) and water (50 ml.), with back washing with small amounts of chloroform. The chloroform solution was dried over sodium sulfate and concentrated under reduced pressure. The crystalline residue, 3,20- diketo 5,6 dichloro 17 a hydroxy 21 acetoxy pregnane, was washed onto a funnel with anhydrous ether. The white fibers of 3,20-diketo-5,6-dichloro-17 e-hydroxy- Zl-acetoXy-pregnane decomposed at 198-198.5 C.

EXAMPLE 12 A -3,20-diketo-1 7a-hydroxy-21-acetoxy-pregnane 0.5 g. of 3,20-diketo-5,6-dichloro-17ahydroxy-2l-acetoxy-pregnane was dissolved in warm acetic acid (25 ml.) and with stirring was treated over a period of one hour with zinc dust in five portions of 0.1 g. each at progressively increasing temperatures beginning at 40 C. and ending at 80 C. The mixture was filtered and the cake was washed with acetic acid (10 ml.). The filtrate plus wash was diluted slowly with water ml.) and the suspension was chilled for two hours and filtered. The crystals of A -3,20-diketo-l7a-hydroxy-2l-acetoxy-pregnene (Substance S acetate) were washed with water, dried, and recrystallized from acetone.

EXAMPLE 13 A -3,20-diket0-1 7a-hydroxy-21-acetoxy-pregnene A solution of chromous chloride was prepared by treating a solution of chromic chloride (10 g.) in water (40 ml.) containing concentrated hydrochloric acid (4 ml.) with amalgamated zinc dust (20 g.). The solution of chromous chloride was added to a solution of 3,20-diketo- 5,6-dichloro-17u-hydroxy-2l-acetoxy-pregnane (0.50 g.) in acetone (70 ml.) under a carbon dioxide atmosphere. After a few minutes standing at room temperature crystallization occurred. After one hour water was added and the acetone was removed under reduced pressure. The resulting suspension of crystals was chilled and the crystals were collected, washed with water and dried, giving material melting at 218-225 C. This material showed A max. at 2410 with E% of 267, indicating that about 60% of the 11 -3,2O-diketo-17a-hydroxy-2l-acetoxy-pregnene had already been isomerized to Substance S acetate. Complete isomerization was effected by adding to a hot solution of 0.100 g. of the material in acetone (5 ml.) 6 drops of a solution prepared from 0.3 ml. of concentrated sulfuric acid and 10 ml. of acetone. The solution was boiled for six minutes then cooled and the resulting crystals of Substance S acetate were separated. After drying, the product melted at 234236 C., showed A max. at 2400 A. (E% =419) and its infrared spectrum was identical with the infrared spectrum of an authentic specimen of Substance S acetate.

EXAMPLE 14 3 ,20-diket0-5 ,6 -dichl0r0-1 7zx-hydroxy-pregnane Ten grams of 3-hydroxy-5,6-dichloro-17a-hydroxy-20- keto-pregnane was dissolved in glacial acetic acid (350 ml.). Water (35 ml.) was added and the solution was cooled to 4 C. A solution of 2.48 g. of chromium trioxide in 2.48 ml. of water and 22 ml. of acetic acid was added over four minutes with stirring and in an ice bath. Concentrated sulfuric acid (1.4 ml.) was added over five minutes and then the reaction mixture was stirred further in the ice bath for 100 minutes. The mixture was then shaken with chloroform (437 ml.) and water (700 ml.). The aqueous layer was separated and re-extracted with chloroform (100 ml.). The combined chloroform layer was washed with water (500 ml.), 2.5% aqueous sodium bicarbonate solution (2 x 500 ml.) and water (500 ml.). The chloroform solution was dried with sodium sulfate and concentrated under reduced pressure with a minimum of heat. The residue was slurried with ether, filtered, washed with ether and dried. The 17a-hydroxy-5,6-dichloro-3,ZO-diketo-pregnane so obtained decomposed at -160 C.

EXAMPLE 15 A -3,20-diket0-1 7ot-hydr0xy-pregnene To a solution of 6.13 g. of 17a-hydroxy-5,6-dichloro- 3,20-diketo-pregnane (prepared as in Example 14) in acetone (800 ml.) was added 500 ml. of chromous chloride prepared as described in Example 13. The solution was concentrated under reduced pressure and the resulting suspension of crystals was chilled and filtered and the product was washed with water. When dry it melted at -205" C.

T o a solution of 4.43 g. of the above material in acetone (100 ml.) at the boiling point was added 5.31 ml. of a solution of concentrated sulfuric acid (0.3 ml.) in acetone (10 ml.). The solution was boiled six minutes, 60 ml. of acetone distilling off in the process. The solution was chilled and the resulting crystals of 17 a-hydroxyprogester' 1 I one were collected, washed with cold acetone and dried. The product, A 3,20 diketo 17a hydroxy pregnene, melted at 210215 C.

EXAMPLE 16 3-acetoxy-5 6 -dichloro-1 7 a-h ydroxy-ZO-keto-Pregn one A mixture of 5.00 g. of 3,20-diacetoxy-5,6-dichloro- 17,20-epoxy pregnane, 30 cc. of methanol and 0.1 cc. of

claims, they are to'be considered as part of our invention.

We claim: 1. A compound of the formula:

wherein R represents a lower fatty acid radical.

The mixture was then.

2. A -3,ZO-diacetoxy-S,fi-dichloro-pregnene. 3. A compound of the formula:

wherein R is a lower fatty acid radical.

Ref

iacetoxy-5,6-dichloro-l7,20-epoxy-pregnane. dihydroxy-S ,6 -dichloro-20-keto-pregnane.

erences Cited in the file of this patent UNITED STATES PATENTS Butenandt Dec. 9, 1941 Butenandt Mar. 16, 1943 Ruzicka Dec. 19, 1944 Mofiett May 6, 1952 Kaufmann May 13, 1952 Cutler et al. Mar. 26, 1957 FOREIGN PATENTS Great Britain Apr. 3, 1936 OTHER REFERENCES Heer: Helv. Chim. Acta 34, 359-372 (1941). 

1. A COMPOUND OF THE FORMULA: 